Callus Induction and Organogenesis Capacity from Lamina Explant of Petunia × hybrida F1 Induced by BA and NAA

Document Type: Original Article


Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran


Plant growth regulators (PGRs) have an important role in callus induction and organogenesis of plant explants cultured in vitro conditions. Callus has a proper potential for plant regeneration through indirect organogenesis and embryogenesis as well suspension culture, genetic transformation and production of secondary metabolites. In current experiment, leaf explants of Petunia × hybrida F1 were cultured on basal Murashige and Skoog (MS) medium supplemented with different concentrations of N6-benzyladenine (BA) (0.25, 0.50 and 1.00 mg l1) and ɑ-naphthalene acetic acid (NAA) (0.10, 0.20 and 0.30 mg l1). The maximum fresh weight (5.16 g), dry weight (0.31 g) and volume of callus (24.50 cm3) was obtained in the medium containing 1.00 mg l1 BA in combination with 0.30 mg l1 NAA. Leaf explants did not produce callus in medium without PGRs. Plantlets were produced on all callus grown on the media containing PGRs. The most plantlets number was produced on callus induced on medium enriched with 1.00 mg l1 BA in combination with 0.30 mg l1 NAA. The plantlets regenerated in vitro with well-developed shoots and roots were successfully established in pots containing peat and perlite (1:1) and grown in a greenhouse within 4 weeks with a 100% survival rate. The regenerated plants were morphologically identical with mother plants and did not show any detectable phenotypic variation.

Graphical Abstract

Callus Induction and Organogenesis Capacity from Lamina Explant of Petunia × hybrida F1 Induced by BA and NAA


This paper presents a simple manner for obtaining callus from lamina explant.

All media containing plant growth regulators induced callus formation.

Plantlets were produced from callus during indirect organogenesis.


Abu-Qaoud, H. 2012. Improving adventitious shoot regeneration from cultured leaf explants of Petunia hybrida using thidiazuron. African Journal of Biotechnology, 11 (51): 11230-11235.

Abu-Qaoud, H., Abu-Rayya, H. and Yaish, S. 2010. In vitro regeneration and somaclonal variation of Petunia hybrida. Journal of Fruit and Ornamental Plant Research, 18 (1): 71-81.

Al-Mallah, Sh. and Salih, S.M. 2006. A protocol for shoot regeneration from leaves petioles tissue culture of neem trees (Melia azedarach). Mesopotamia Journal of Agriculture, 34 (1): 1-8.

Clapa, D. and Cantor, M. 2006. Plant regeneration from stem cuttings of Petunia hybrida. Bulletin of University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca. Horticulture, 63 (1-2): 45-49.

George, E.F., Hall, M.A. and Klerk, J.D. 2008. Plant Propagation by Tissue Culture. The Background, Springer, 1: 65-75.

Guang-rong, C., Ji-tian, L. and Hai, C. 2004. Study on tissue culture of petunia hybrid. Special Wild Economy and Animal Plant Research, 2004-2003.

Jain, S.M. and Häggman, H. 2007. Protocols for Micropropagation of Woody Trees and Fruits. Springer Publication.

Jain, S.M. and Ochatt, S.J. 2010. Protocols for In Vitro Propagation of Ornamental Plants. Springer Protocols, Humana Press.

Maberly, D.J. 1990. The Plant Book. A portable dictionary of the higher plants. Cambridge University Press, Cambridge, U.K.

Mihaljevi, S., Bjedov, I. and Kova, M. 2002. Effect of explant source and growth regulators on in vitro callus growth of Taxus baccata L. Washingtonii Food Technology and Biotechnology, 4: 299-303.

Murashige, T. and Skoog, T. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plant, 154: 473-497.

Qu, Y.H., Lin, C., Zhou, W., Li, Y., Chen, B. and Chen, G.Q. 2009. Effect of CO2 concentration and moisture content of sugar-free media on the tissue–cultured plantlets in a large growth chamber. Communications in Nonlinear Science and Numerical Simulation, 14 (1): 322-330.

Praveen, N., Naik, P.M., Manohar, S.H., Nayeem, A. and Murthy, H.N. 2010. In vitro regeneration of brahmi shoots using semisolid and liquid cultures and quantitative analysis of bacoside A. Acta Physiology Plant, 31: 723–728.

Reuveni, M. and Evenor, D. 2007. On the effect of light on shoot regeneration in Petunia. Plant Cell Tissue and Organ Culture, 89: 49-54.

Rui-yue, Z. 2007. Tissue culture and rapid propagation of Petunia hybrida. Journal of Anhui Agricultural Science, 2007-2010.

Sriskandaraja, S., Prinson, E., Motyka, V., Dobrev, P.I. and Serek, M. 2006. Regenerative capacity of Cacti Schlumbergera and Rhipaslidopsis in relation to endogenous phytohormones, cytokinin oxidase/dehydrogenase, and peroxidase activities. Journal of Plant Growth Regulation, 25 (1): 79- 88.

Subotic, A., Jevremovic, S., Cingel, A. and Milosevic, S. 2008. Effect of urea-type cytokinins on axillary shoot regeneration of Impatiens walleriana L. Biotechnology and Biotechnological Equipment, 22 (3): 817-819.

Thirukkumaran, G., Ntui, V.O., Sherkhan, R. and Mii, M. 2009. Thidiazuron: an efficient plant growth regulator for enhancing Agrobacterium mediated transformation in Petunia hybrid. Plant Cell Tissue and Organ Culture, 99 (1): 109-115.

Xian-Chun, Z. 2010. Effect of the plant hormones ratio on tissue culture of Petunia hybrida. Journal of Anhui Agricultural Science, 17-20.